I was recently browsing across the Paul N. Hengen's excellent
Molecular Biology Homepage (http://www-lmmb.ncifcrf.gov/~pnh/),
and I found a technique used for storing RNA that seems to be greatly
preventative towards degradation.
This technique seems too good to be true; extremely inhibitory to
RNase activity and no apparent problems with its use, and it.
The only thing that make me a cautious about this technique is that
not one lab that I know uses it; they all suspend RNA in either DEPC
or ethanol.
Has anyone had experience with using the following RNA storage
technique, and is there something that I should know about before I
start resuspending my samples this way?
Appreciate any insights or experience that people have!
Jason
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U.S. Dept Commerce/NOAA/NMFS/NWFSC/Molecular Biology Protocols
Solublization of RNA in Formamide
contributed by James McCaughern-Carucci, Yale University
Resuspending RNA in Formamide (as reported by Chomczynski et al.
Nucleic Acids Research) has several benefits over storage in water or
ethanol. First, formamide will protect RNA from RNases. The RNA is
also extremely stable, allowing it to be stored overnight at 4C, or
indefinitely at -20C. Samples can even be left at room temperature
overnight without fear of degradation. Samples can be made to be
highly concentrated, up to 4 mg/ml. Finally, with concentrated
samples, no drying of your RNA is necessary for running Northerns,
RT-PCR, or RNase Protection, saving time and potential degradation.